GSM7892771: melanocyes, H3K27ac ChIPseq, rep1; Columba livia; ChIP-Seq
Melanocytes were fixed with 1% formaldehyde for 15 min and quenched with 0.125 M glycine. Chromatin was isolated by adding lysis buffer, followed by disruption with a Dounce homogenizer. Lysates were sonicated and the DNA sheared to an average length of 300-500 bp with Active Motif's EpiShear probe sonicator (cat# 53051). Genomic DNA (Input) was prepared by treating aliquots of chromatin with RNase, proteinase K and heat for de-crosslinking, followed by SPRI beads clean up (Beckman Coulter) and quantitation by Clariostar (BMG Labtech). Extrapolation to the original chromatin volume allowed determination of the total chromatin yield. Illumina sequencing libraries (a custom type, using the same paired read adapter oligonucleotides described by Bentley et al., (2008)) were prepared from the ChIP and Input DNAs on an automated system (Apollo 342, Wafergen Biosystems/Takara). After a final PCR amplification step, the resulting DNA libraries were quantified and sequenced on Illumina's  NovaSeq 6000 (75 nt reads, single end). Illumina library: Bentley D.R. et al., Nature. 2008 November 6; 456(7218): 53–59. doi:10.1038/nature07517, PMCID: PMC2581791